Our Technologies

Electroporation is a widely used and highly efficient transfection technique. It employs brief electrical pulses to generate transient pores in cell membranes, enabling the delivery of diverse molecules—including DNA, RNA, mRNA, ribonucleoproteins (RNPs), and proteins—into virtually any cell type.

The conventional method of delivering materials into the cell nucleus, pronuclear injection, involves using a very thin glass needle to inject materials into the nucleus. However, using this method, the amount of materials being injected cannot be controlled well while there are also chances that the glass needle may damage the DNA in the pronucleus and the membrane of the zygote. The survival rate after injection and birth rate after injected embryos being transferred to recipient mouse are greatly reduced because of the above limitations. At least 2-3 generations (about 9-12 months) of breeding is needed to get the desired experimental colonies. 

Electroporation employs a straightforward procedure: cells and target molecules are combined in a conductive buffer within an electrical circuit. A precisely calibrated, microsecond to millisecond pulse is then applied. This pulse transiently disrupts the cell membrane's phospholipid bilayer, creating temporary pores. Concurrently, the resulting electric field drives charged molecules (e.g., DNA) across the membrane through these pores via an electrophoretic effect.  Using GTL’s optimized protocol, we are able to deliver materials into the pronucleus of both male and female efficiently while doing no damage to embryonic development. The amount of time needed to yield the desired experimental mouse is greatly reduced to one generation (about 3 months).  

The technique offers a combination of efficiency and practicality: it is relatively simple to execute, yields highly reproducible results, and enables the rapid transfection of a large number of cells. Importantly, it requires no vector and is less dependent on cell type, which makes it particularly suitable for challenging-to-transfect cell lines.

Electroporation can deliver the desired experimental animal in a shorter period of time while also having a lower cost compared to the traditional pronuclear microinjection method.

 The results of knockout via electroporation targeting 19 genes showed an insertion/deletion (Indel) rate of 93.2% and a homozygous knockout (hom KO) rate of 85.6%. In contrast, the knockout via pronuclear injection targeting 2 genes resulted in an Indel rate of 42.6%, but no homozygous knockout individuals were detected. 

ES cell injection is one of the most widely used methods for creating gene-targeted mice, including knock-outs and knock-ins. This method involves injecting genetically modified embryonic stem cells into mouse blastocysts. In GTL’s approach, ES cells are introduced into 8-cell stage embryos using a finely pulled micro glass needle. Following the injection, the resulting blastocysts are surgically transferred into the uteri of pseudopregnant surrogate mothers for generation of genetically modified mouse.

Using GTL's ES cell injection method, we are able to produce fully ES cell-derived pups in the F0 generation, greatly reducing the time needed to obtain experimental mouse.

The efficiency of genome editing is heavily dependent on the sequence of guide RNA and cell types. Although there are existing software that can precisely predict the editing results in somatic cells, however, there is no software that can predict the editing results of an embryo. GTL has developed and constantly improving two new software using published somatic editing data that are comparable with commercial prediction software.